HEMOSTATIC PATCH

Abstract: A hemostatic patch that is advantageously safe and inexpensive is created that comprises a biodegradable matrix such as an absorbable gelating sponge, and an effective amount of epsilon aminocaproic acid for inhibiting fibrinolysis. Epsilon aminocaproic acid is a hemostatic agent that inhibits fibrinolysis, accelerates the activity of thrombin and possesses antibacterial properties. A patch containing EACA and calcium chloride is effective for decreasing bleeding of parenchymal organs, as well as for topical use particularly in a bandage form. The patch further can contain thrombin, RGD and protamine sulfate.

Description: HEMOSTATIC PATCH Background of the Invention A hemorrhage of a blood vessel, body tissue, organ or bone can result in blood loss leading to hypovolemic shock and death. In hemophiliacs and patients receiving anticoagulant medication, such as often prescribed post- operatively for heart surgery, the problem of rapid blood loss is even more acute. Attempts have been made to devise a fast, effective and inexpensive method for curbing blood loss, including pastes containing coagulation-enhancing factors. One such coagulation enhancing substance employed to assist a cessation of bleeding or "hemostasis" is human fibrinogen, most commonly employed as a "fibrin glue". Fibrin glue is composed of a mixture of human fibrinogen and bovine thrombin. It is sold as a kit containing separate vials of fibrinogen and thrombin solutions. These solutions are mixed together and applied to the wound in various ways, including as a paste, as a spray or on a patch. Fibrin glue, however, is an inconsistent and ineffective therapy for hemostasis. The mixing, soaking, and coating of a patch with fibrin glue requires time- consuming and cumbersome procedures. Each of the preparation steps introduces potential errors and thus their efficacy varies with the experience of operating room personnel. Moreover, during the preparation of such solution, further hemorrhage occurs and the solutions are washed away by intense bleeding. Despite the headway made in fibrinogen compositions and surgical techniques, these pitfalls in achieving hemostasis underscore the need for development of a suitable product. An improvement over fibrin glue, marketed in Europe consists of a biodegradable collagen patch onto which is impregnated bovine thrombin, aprotinin and human fibrinogen (the "TAF" patch) . An example of a TAF patch is the TachoComb® patch marketed in Europe by Hafslund Nycomed Pharma, DE. The patch also contains calcium chloride to enhance coagulation. In use, this patch is removed from its package, dipped into saline solution and applied to the bleeding organ with light pressure for at least five minutes. When the bleeding has stopped, the patch is left in place by the surgeon and the cavity closed. A major drawback to the use of fibrin glue and the TAF patch is that both contain human fibrinogen, a protein purified from human blood. Because of the high risk of HIV and hepatitis viral contamination, the Food and Drug Administration revoked the use of human fibrinogen in the United States in 1978. In addition to the safety concerns, human fibrinogen purified from human plasma is very expensive. A TAF patch also requires refrigeration in order to stabilize the coagulation-enhancing agents contained in the patch. This requirement prohibits certain field applications of the patch, where refrigeration facilities are unavailable. Another problem with a TAF patch that surgeons cite is its inflexibility, that is, the patch does not conform easily to the shape of the body surface to which it is applied. A hemorrhage of a parenchymal organ, such as the spleen, liver, lung or pancreas, which can result from trauma or surgery, is particularly difficult to treat. Parenchymal organs are difficult to ligate because the tissue is easily torn, pulverized or crumbled. As a result, surgeons often resort to the use of electrocautery, which can lead to further destruction of the patient's tissues. Thus, an effective hemostatic patch is desired which is safe from deadly viral contamination and even stops bleeding in the problematic hemorrhages of parenchymal organs. A patch is further desired that is inexpensive, easy to use and that molds easily to body contours. Also a need exists for a patch that withstands elevated temperatures without requiring refrigeration and retains hemostatic efficacy. Summary of the Invention According to the present invention, an effective hemostatic patch is produced comprising a matrix and at least one hemostatic agent, epsilon aminocaproic acid. The patch does not require as an ingredient any exogenous human protein, such as fibrinogen, which thereby avoids introduction of unsafe contaminating viruses. The present hemostatic patch is inexpensive, easy to use, thermally stable, antibacterial, and also can be provided in bandage form for topical use. Brief Description of the Drawings Figure 1 displays a control experiment showing the thrombin activation at 37°C and physiological pH. Figure 2 demonstrates the effects of pH on thrombin activation at 37°C. Figure 3A and Figure 3B each show inhibition by EACA of Staph. aureuε growth in the presence of various concentrations of EACA. Figures 4A and Figure 4B each show inhibition by EACA of E . coli growth in the presence of various concentrations of EACA. Figure 5 demonstrates a Tachocomb® ("T") patch, two Hemarrest™ patches, and blank "B" gelatin foam patch, which are compared in Example 3. Figure 6 shows photographically a liver lesion study in which the TachoComb® patch (middle) is compared with two GT(Ca++)E patches according to the present invention. Figure 7 shows a photograph of the same liver shown in Figure 6 with the patches removed from the lesions. Figure 8 shows photographically a Hemarrest™ patch of the invention, containing 100mg/cm 2 epsilon aminocaproic acid, as applied to a kidney. Figure 9 shows photographically the opposite pole of the same kidney of Figure 8, with a TachoComb® patch applied. Figure 10 shows photographically a comparison between the GE and GTE patches of the invention. Figure 11 shows photographically a comparison between matrices of calcium/sodium alginate, collagen and gelfoam. Patches CVAT(Ca++)EF, "Cal-Alg"; CVCT(Ca-H-)EF, "Collagen"; and GT(CA++)EF, "Gelfoam" of the present invention are shown. Figure 12A shows photographically a control set of unused GT(Ca++)EF, CT(Ca++)EF and GT(Ca++)EFP patches of the present invention, while Figure 12B shows these patches as applied to the spleen. Figure 13A shows a side view of a bandage embodiment of the invention, including a patch of the invention, while Figure 13B shows an elevation view of the bandage. Detailed Description of the Preferred Embodiments According to the present invention, a hemostatic patch is provided that comprises a shaped structural element that is a biodegradable matrix, such as absorbable gelatin sponge or calcium alginate, to which is applied a hemostatic agent that contains epsilon aminocaproic acid, "EACA." EACA is an inhibitor of clot degradation. In the body, clot formation and clot breakdown are competing processes. EACA inhibits the production of plasmin, an enzyme that degrades clots. Plas in degrades clots by solubilizing fibrin, an important component of clots, in a process called fibrinolysis. By inhibiting the formation of plasmin which breaks down clots, EACA inhibits fibrinolysis and drives the reaction conditions at the patch/biological interface in favor of clot formation. A hemostatic patch according to the invention thus comprises an amount of EACA effective for inhibiting fibrinolysis. In a preferred embodiment, a hemostatic patch containing EACA is provided that comprises a flat flexible matrix which is biodegradable and absorbable. Further the matrix has an uncompressed thickness of about 4-10 mm, or a compressed thickness of about 2-10 mm, advantageously 2-5mm, and a length of about 1-2 to 20 cm, more preferably between 4-10 cm. The matrix, of course may be cut to a desired length. Preferably, EACA is applied onto only one side of the matrix, the wound- contacting surface, in amounts of 10-100 mg/cm 2 , more advantageously 60-70 mg/cm 2 . Optionally, one or more additives, including a calcium ion source, RGD peptide, RGDS peptide, protamine sulfate and buffer, are applied to that surface. Further, about 1-1000 IU/cm 2 , advantageously 1-100 IU/cm 2 , and more advantageously 1-4 IU/cm 2 , is applied to the wound-contacting surface. A preferred amount applied in this context is 1.25 IU/cm 2 of thrombin. By applying hemostatic agent(s) and additive(ε) as defined herein to a single side of a flat patch, namely, the wound-contacting surface, the patch is much less susceptible to forming undesirable adhesions between tissue surfaces. Such adhesions are particularly problematic with hemostatic patches designed for internal/surgical use. Abnormal adhesions result when non-wounded, normal tissue in the surrounding vicinity of the wound comes into contact with the patch covering the wounded surface, and then stably binds or "heals" to the wound abnormally. For example, an ulcerative intestine to which a competitor's patch is applied may abnormally adhere to a normal segment of intestine which overlies it. As a result, the normal mobility of that intestinal segment decreases. Histological sections of a preferred patch, such as the "Hemarrest™" patch as defined herein, showed that when left for sufficient period of time in situ to permit hemostasis, advantageously adhesions did not form between non-wounded, normal tissue and the patch. It is contemplated that other embodiments of the patch, such as listed in Table 1 below, also would avoid adhesions as well. The avoidance of abnormal adhesions demonstrated by a patch containing EACA is an important feature, because, surprisingly, it has been discovered that EACA functions as a hemostatic agent in a patch in a manner that approximates the effectiveness of fibrinogen, a coagulation factor. Fibrinogen, in solution, converts to fibrin in the presence of thrombin, and is an active ingredient found in other hemostatic patches. EACA, however, is devoid of the hazards that accompany use of fibrinogen. Moreover, according to the present invention, it has been determined, surprisingly that EACA in the matrix of a patch provides an alkaline environment that accelerates the activation of thrombin. In comparison with thrombin activation measured in the absence of EACA (Figure 1, closed boxes) , EACA greatly increases thrombin's activity (Figure 2) . This phenomenon holds true whether the EACA acts on thrombin present in the blood endogenously or on thrombin that is supplied externally in a patch. Thus, it has been discovered that a patch comprising EACA exerts a dual hemostatic action by (1) slowing clot degradation by inhibiting plasmin formation and (2) accelerating clot formation by activating thrombin. Therefore, a method is provided for accelerating the activity of thrombin by increasing the pH of the local environment of a patch according to the invention. Such a patch comprises a matrix and a "thrombin enhancing compound" capable of raising the pH in a solution in the local environment of the patch sufficient to increase the activation of thrombin. Such a compound is capable of raising the pH of the local environment to a pH in the range of 7.0-9.0 inclusive, more advantageously between pH 7.02-8.02 inclusively, and even further advantageously, between pH 7.62-8.02, inclusively. According to the present invention, an alkaline solution is created in the local environment of the patch as the thrombin enhancing compound εolubilizes upon its contact with blood. Then, thrombin present in the blood and, optionally, thrombin provided as an exogenous ingredient of the patch mixes with the alkaline solution in the local environment of a patch and thereby is activated. Advantageouεly, the thro bin-enhancing compound provided in the patch for increaεing pH is EACA. A "sterile buffer" which is pharmaceutically acceptable and capable of buffering the local pH in the patch to alkaline conditions, (i.e., between a pH of 7.0-9.0, more advantageously pH 7.02-8.02, and even further advantageously, 7.62-8.02), is suitable as a thrombin- enhancing compound, as well. For example, Tris buffer is an effective thrombin-enhancing sterile buffer, as shown in Figure 2, open diamond-shaped graphical plot. Other sterile buffers that buffer the pH in this range are contemplated, such as Hepes buffer, for example. Accordingly, in a more advantageous patch, EACA and Triε (or other) buffer both are provided in the matrix of the patch. Yet another surprising advantage of EACA has been discovered. EACA possesses antibacterial properties. According to the present invention, it has been demonstrated that EACA exerts dose-dependent inhibition of both S. aureus and E . coli growth (Figures 3A, 3B and 4A, 4B, respectively) . Therefore, the EACA/matrix patch according to the present invention is very desirable for its antibacterial effects on microorganisms present at the wound site where a patch is applied. Another advantage of EACA is that it contains no foreign peptides of animal origin. For example, a non- human fibrinogen hemostatic agent in some humans will trigger an immune response or allergic-like reaction. Thus, a patch according to the invention can contain as a sole hemostatic agent EACA dispersed within a matrix or applied to a surface of a matrix in an amount effective for inhibiting fibrinolysis and thereby stimulating clot formation. A biodegradable "matrix" as referred to herein, and as employed in any of the present embodiments of the invention, is selected from, but not limited to, the group consisting of absorbable gelatin sponge, calcium alginate, calcium/sodium alginate, collagen, and oxidized regenerated cellulose. A matrix of other forms of collagen, such as crosslinked collagen, esterified collagen or chemically modified collagen as taught by U.S. Patent No. 4,390,519 to Sawyer, and other conventional matrices utilized in hemostatic patches, are contemplated for use with EACA according to the present invention. Four matriceε that are advantageous for use with EACA include absorbable gelatin sponge, calcium alginate, calcium/sodium alginate, and collagen. A first embodiment of the invention therefore provides a patch comprising a matrix of absorbable gelatin sponge "G" and a hemostatic agent, EACA "E." This embodiment, "GE", preferably also can contain calcium, "G(Ca++)E." Advantageously, the GE or G(Ca++)E patch need not contain thrombin or fibrinogen to function effectively. As a result, both GE and G(Ca++)E, have good thermal stability and can be stored for months to a few years without refrigeration and losing effectiveness. The GE and G(Ca++)E patches are useful for field and emergency use, since each may be stored in a ready-to-use state for a lengthy period, even in absence of refrigeration. Both also are much less expensive to make a patches which contain fibrinogen. The many representative embodiments of the present invention are referred to herein most easily by acronyms, e.g., GE. These acronyms are indicative of the individual components (Table 1) found in the patches created in accordance with the invention. Table 1 . PATCH COMPONENT CODES: G = gelatin foam patch alone, e . g . , Gelfoam® CA = calcium alginate CVA = calcium/sodium alginate, e.g., Kaltostat® C or CVC = collagen or collagen(Helistat®) , respectively E = EACA (Ca++) = calcium T = thrombin R = RGD peptide P = protamine sulfate F = Fibrinogen (f) = freshly applied compound (Example 7) GT(Ca++)E= "Hemarrest™" patch In other embodiments, a GE or G(Ca++)E patch further comprises an effective amount of thrombin for stimulating hemostasis and thus is designated as "GTE" or "GT(Ca++)E." A thrombin molecule is most stable at temperatures between 2-8°C. However, these patches can be stored for a limited period of time at room temperature. In fact, because addition of thrombin enhances the GE and G(Ca++)E patches' effectivenesε, these patches are very useful outside the clinic for field use, such as for emergency or military purposes. Although it is understood that exposure to extreme environmental conditions may render thrombin present in the patch partially or totally inactive, the activity of the remaining GE or G(Ca++)E patch would not be substantially affected. The GTE and G(Ca++)E patches further can contain fibrinogen and are designated as GTEF and GT(Ca++)EF, respectively. In the GE and GE(Ca++) patches, and all patches described herein that employ an absorbable gelatin sponge usp as a matrix, the matrix is advantageously a flat layer of gelatin foam, more advantageously, Gelfoam®, and even more advantageously, compressed gelatin foam or compressed GelFoam®. The effectiveness of patches of the present invention in promoting clot formation is enhanced by the lattice structure of the gelatin foam, which promotes enzyme substrate interactions. In particular, the gelatin foam structure enhances contact between thrombin provided exogenously in the patch with endogenouε fibrinogen preεent in the blood exuding from the wound. Additional hemoεtatic agents can be applied to the GE patch in amounts effective for εtimulating hemoεtasis, including, but not limited to: thrombin "T", an enzyme which convertε fibrinogen to fibrin; calcium, sodium, magnesium or other ionε that εtimulate hemostasis; and optionally, fibrinogen, "F". In terms of ion additives, calcium chloride is generally a preferred additive for introducing a calcium ion into the patch, particularly where that patch does not contain exogenous fibrinogen. In a patch containing fibrinogen, however, calcium chloride and sodium chloride function about equally as well as hemostatic additives in promoting clot formation. "EACA analogs," or compounds that possess a similar hemostatic activity and a chemical structure to that of EACA, can be used instead of, or in addition to, EACA in a patch according to the invention. Possible EACA analogs contemplated for addition to a matrix include EACA derivatives having bioisosteric functional groups. EACA'ε carboxylic acid group can be substituted, for example, by sulfonic or sulfinic acid (-S0 2 H and -S0 3 H) or phosphonic acid groups. Examples of analogs include, but are not limited, to 5-aminopentanoic a acid, 7- aminoheptanoic acid, 8-aminooctanoic acid, provided that these compounds exert a hemostatic activity. The molecules "thrombin" and "fibrinogen" as defined herein are meant to include natural thrombin and fibrinogen molecules derived from an animal or human origin, a synthetic form or a reco binant form of the molecules, including functionally active analogs that effectively maintain the enzyme's clot promoting activity in an animal or human. The species of animal from which - li ¬ the molecule is derived can vary and dependε on the intended use of the patch. For example, a patch intended for human use for safety reasons contains non-human thrombin and non-human fibrinogen, and preferred in this context iε bovine thrombin and bovine fibrinogen. By avoiding use of human fibrinogen, risks associated with viral contamination of purified blood products (particularly with fibrinogen) are minimized. Indeed, the ingredients EACA, thrombin and GelFoam® all are approved by the U.S. Food and Drug Administration for human use. In yet another embodiment, a patch is provided having a matrix composed of calcium-sodium alginate "CVA" or calcium alginate "CA," and a hemostatic layer of EACA "E." It is understood that calcium alginate can be substituted for calcium/sodium alginate in the discussion and examples hereafter, without substantial differences in results. The embodiment, "CVAE", advantageously contains calcium ion and thrombin as well. It also iε less expensive as compared with a patch that containε fibrinogen. Similar to the GE patch, the CAE patch can include additional hemostatic agents including, but not limited to, thrombin, calcium or sodium or other ions in amounts that are effective to εtimulate or accelerate hemostasis. Optionally, fibrinogen can be added in amounts sufficient to stimulate or accelerate hemostasis, as well. These patches further can contain additives as described herein, as well. In another embodiment, an effective amount of the active peptide, RGD, "R" or RGDS effective to stimulate wound healing is added to a patch compriεing GE or CAE, and thus such a patch is designated as GER or CAER. The tripeptide RGD is composed of arginine, glycine and aspartic acid, and optionally serine "RGDS," and iε the active εite of fibrinogen and fibronectin. RGD accelerates wound healing and is believed to stimulate fibroblast migration. The RGD additive is also much less expensive than fibrinogen. RGD can be synthesized easily using conventional solid phase chemistry at a fraction of the cost of obtaining fibrinogen, which currently must be obtained by purification from a natural εource. In yet another embodiment, an amount of the agent protamine εulfate "P" effective to neutralize heparin present in the local environment of the patch is added to any of the aforementioned patches comprising EACA and a matrix. Protamine sulfate neutralizes heparin or vitamin K antagonists that are present in the blood of certain patients or animals being treated with a hemostatic patch. A patch comprising GEP or CAEP, for example, iε prescribed for persons undergoing parenteral therapy with heparin. In particular, a patch that further containε thrombin would be effective in patients taking dicumarol. A patch containing protamine sulfate is preferably stored at refrigerated temperatures of 2-8 degrees Celsius to maintain the activity of protamine sulfate. An additional advantage of the patches according to the present invention is that the matrices, such as absorbable gelatin sponge or calcium alginate, and the hemostatic agents, especially EACA and thrombin, and the additive, RGD, all are relatively inexpensive. It iε estimated that production of a "standard-size" rectangular patch of about 9.5 x 4.8 cm, having a thickneεs of about 2.5 mm would cost substantially less than a TAF patch of the εa e εize. Patches according to the present invention exhibited efficacy in inducing hemostasis in freely bleeding lesions of the spleen, liver and kidney of an anesthetized pig- Surgical lesions induced in parenchymal organs of pigs provide a good model system for hemostasis in the analogous human organs as evidenced by preclinical studies performed on pigs and dogs for the TachoComb® patch. Schiele et al . , Clinical Materials 9:169 at page 172 (1992). See also, SWINE AS MODELS IN BIOMEDICAL RESEARCH, Swindle, M. , Iowa State Univ. Press (1992) . Indeed, surprisingly, patches according to the present invention performed better than TachoComb® in the liver, while in the kidneys, the patch containing a matrix of GelFoam®, thrombin and lOOmg/cm 2 EACA performed equally as well as the TachoComb® patch. The results of that comparative experiment are presented in Example 3 herein. Another important advantage of the present invention is its flexibility, that is, a patch is provided that easily conformε to the contourε of an organ or biological surface, making the manipulation of applying the patch quicker to perform. Aε a reεult, there iε less overall blood loss to the patient and less time is spent in surgery. A hemostatic patch according to the present invention in employed by applying a "wound-contacting" surface of the patch, a surface intended to contact the wound and containing hemostatic agent(s) and optionally additiveε, to a bleeding wound. Then, the patch is maintained in contact with the wound for a period of time sufficient for clotting to occur at the interface between the hemostatic patch and the wound and for bleeding to be substantially arrested. Preferably the patch iε maintained in contact with the wound εurface for a period of about 3-20 minutes, advantageously 3-10 minutes, and more advantageously, 3-5 minutes. Where EACA, thrombin, and calcium chloride all are preεent on/in the matrix, the time period iε preferably about 5 minutes. The patch is held in place againεt the biological εurface preferably with light pressure, preferably by means of a sterile saline soaked sponge. Alternatively, the patch may be held in place simply by applying pressure to the patch by means of a gauze or other dry sterile material. Depending on the location of the wound, a bandage, including an elasticized bandage, can be wrapped around the patch so as to provide light presεure on the wound εite. In addition to inducing hemostasis, a patch according to the present invention is useful for hermetically sealing body tisεue. For example, when air leaks from a wound in the lungs, a patch is applied to the surface surrounding the wound, held in place with light presεure for a period of time adequate to induce hemostasis, as discussed above. During that time, in addition to hemostaεiε, a hermetic εeal formε. Prior to applying the patch, it iε preferable to εoak the patch in εterile εaline solution. Such a step is not required, however. Use of a hemostatic patch according to the invention, without firεt εoaking in εaline εolution permits quick and simple application of the patch in field situationε, εuch aε may be encountered by an emergency medical technician or a military healthcare worker. In one embodiment, the patch is contained within a sealed sterile package which facilitates removal of the patch without contamination. Such a package for example, can be an aluminum foil pouch or other conventional material that is easily sterilized. Radiation, advantageously gamma radiation, is applied to sterilize the patch and packaging material together. In another embodiment, a container having dual compartments is provided. A firεt compartment containε distilled water, sterile saline or a sterile buffer, while the second compartment contains a patch according to the invention. In field use, the patch of the εecond compartment can be readily dipped into an opened first compartment and subεequently applied to the wound. A preferred uεe of a patch according to the preεent invention iε to inhibit or completely stop bleeding of a parenchymal organ, such as the liver, kidney, spleen, pancreas or lungs. Additional uses for such a patch include curbing bleeding of tissues during types of surgery such aε, but not limited to, internal/abdominal, vaεcular (particularly for anastomosis), urological, gynecological (particularly for an episiotomy) , thyroidal, neurological, ENT, tisεue tranεplant uses, and dental surgeries. Another use of a hemostatic patch includes topical treatment, such as for burn or tissue transplants. A patch intended for topical use according to the invention preferably contains additives, such as anti-infection medicaments. Bactericides, fungicideε and wound healing agentε can be added, aε well. Neomycin and bacitracin are examples of certain additives that are incorporated into a patch intended for topical use, in addition to the antibacterial properties of EACA discussed above. A hemostatic patch of the invention also is useful for treating animals, preferably humans or other mammals. Thus, both companion, livestock and wild animals can be treated with a hemostatic patch. A patch in size and shape according to the intended use. Moreover, a standard εize rectangular patch, 9.5 x 4.8 cm, having an uncompreεsed thickness of about 4-10 mm, or a compresεed thickness of about 2-10 mm, advantageously 2-5mm, may be cut to εize with a pair of scissors. One example of an advantageous matrix to which EACA and hemostatic agents and or other additives according to the invention are applied includes gelatin foam, preferably provided in a compressed form. More preferably, a GelFoam® matrix that is compressed to at least one-half its original thicknesε. Also, a patch may be spherically, conically, cuboidally or cylindrically-shaped or prefabricated into small squares, such as for packing into a body cavity. Such an embodiment is useful for example, for a dental cavity resulting from tooth extraction. Additionally, the patch can be configured into a tampon, for example, for epistaxis (profuεely bleeding nostril) or other void. A patch intended for topical applications additionally can be applied with an adhesive tape, as a band-aid form, where the patch is adhered to an adhesive backing. Preferably the adhesive used to secure the patch is porous in areas which contact the skin. An embodiment of the present invention provides thin film wound dressing or bandage for the application to skin εurfaceε to provide a sterile mechanical barrier to all types of infectious agents. Preferably, the "bandage" embodiment, shown in Figure 13, comprises a backing member 10 located contiguous with an exterior surface of the patch 20, which is opposite that of the wound contacting surface 30. In a preferred embodiment, the backing serves to prevent passage of the hemostatic agent(s) , optional additive(s) and other biological exudants through the exterior surface 30, as well providing a barrier to entry of infectious microorganisms into the patch and underlying wound. The backing also provides physical support and protection for the patch and underlying wound, and preferably is fixedly secured to the exterior side of the patch, for example, with a medically acceptible adhesive. Various occlusive and non-occlusive, flexible or non- flexible backing members can be used in the adhesive bandage of the invention. Preferable for use in this context are several polyurethane films that have been specifically adapted for wound dressingε and other medical uses. These films are typically used in thicknesses of less than 2 mm and allow the free diffusion of oxygen, water vapor and other gasseε through their molecular matriceε. In addition, theεe filmε are impermeable to both liquids and all known microbial disease vectors. Other suitable backings can include polyethylene, acrylic, silicone, cellophane, cellulose acetate, ethylcellulose, plaεticized vinylacetate-vinylchloride copolymers, polyethylene terephthalate, nylon, polyethylene, polypropylene, polyvinylidenechloride, paper, cloth, aluminum foil and other conventional backing materials. Preferably, a flexible backing material iε employed to permit the bandage to readily conform to the contour of the patient's body member to which the patch is applied. In providing a bandage according to the invention, a flap 15 extends from the backing member, beyond the region of the patch. In another embodiment, the flap(s) is fixedly secured to patch itself, preferably on the εides of the patch. Advantageously, at least two flaps are provided which extend from the backing member, on opposite sides of the length, or of the width of the patch member. Alternatively, a plurality of contiguous flaps may extend in overhanging fashion outwardly from the backing member beyond the patch, such as shown in Figure 13. Where a flap extendε outwardly from a εingle side of the backing member, the flap preferably is sufficiently long to permit its encircling an appendage, such as a finger, and adhering to the backing member on the opposite side of the patch. A medically acceptable adhesive 25 is applied onto the flap(s) , to permit adherence between the backing member and the patient's skin. Preferably, the flap(s) and the backing member are made of material which permitε the skin to breathe. The backing and flap material can be made of the same or different materials. A removable "pull-tab" 40, comprising a layer of cellophane, plastic or the like, is applied to the flap(s) to prevent it (them) from sticking to other surfaces prior to use. To use the topical adhesive bandage of the invention, the pull-tab layer(s) are readily peeled off just before application of the bandage to the body. Then the bandage is applied with the wound- contacting surface facing downward in direct contact with the wound. The flap(ε) preferably doeε not contact the wound, but instead is applied to a region(s) of normal tissue adjacent the wound to secure the patch member in place. One or more additional layers of wound dresεing material, preferably a layer which aids in absorption of blood or other exudants, can be applied to a patch. Such an additional layer can be made as an integral part of the patch, thereby creating a thicker patch. Alternatively, the layer may be applied as a supplement to the backside (non-wound contacting surface) of a patch according to the invention. Particularly for topical use, the layer(s) can contain superabεorbentε to wick exudant solution from the wound site. It is advised that for patches intended for internal-surgical applications, where an added layer(s) iε integral with the patch, the layer(s) should be both biodegradable and pharmaceutically acceptable. The patch can be designed to facilitate its application to anastomose or fuse ends of a blood vessel or other body lumen having been severed εurgically or otherwise. To apply a patch for anastomosis, a rectangular GETR patch, for example, is wrapped around the external surface of the ends of a Dacron® graft. When the graft is positioned into place, the patch accelerates fibrin growth into the graft to seal the graft in place (hemostatically and hermetically) . A kit is provided that containε a graft and a patch according to the present invention that is designed for fitting with the ends of the graft. Alternatively, a kit iε provided having a patch of the present invention pre- fitted onto at least one end of a graft. Preferably, a wound-contacting surface of the patch iε coated with a color indicator to assiεt the user, such as yellow vitamin B 2 (riboflavin) or a suitable dye, for example, hemin. By color coding the patch, the user knowingly avoids touching or otherwiεe contaminating the wound-contacting εurface of the patch. A patch containing EACA according to the invention iε made by applying to a matrix, an amount of EACA effective for inhibiting fibrinolyεis in the local environment of the matrix. Advantageously, about 10-100 mg/cm 2 of EACA is applied to a wound-contacting surface of the matrix, more advantageously 60-70 mg/cm 2 . EACA, as well as the other hemoεtatic agents or additiveε deεcribed aε componentε of a patch according to the invention, can be applied to the matrix by any of several methods which all would be performed most advantageously under sterile conditions. It is understood that conventional methods of applying the hemostatic agents and additives to a matrix compriεing EACA besides those described herein can be performed as well. Advantageously, EACA iε applied aε a layer to a particular surface or side of the matrix, which surface is then designated as the wound-contacting surface. This can be accomplished by εpraying EACA in powder form onto the patch. Alternatively, a εolution of EACA can be coated onto a matrix and dried by lyophilization or by conventional means. In another method of applying EACA, a matrix is dipped completely or partially into a sterile solution of EACA such that a sufficient amount of EACA accumulates within the matrix effective to inhibit fibrinolysis in a mammal, such that similar effectiveness to the Hemarrest patch is demonstrated. In making a patch intended for internal uεe, the matrix is preferably not dipped completely or otherwise saturated with EACA; rather, the EACA is applied to a single side by any of the alternate ways described above. After application of EACA to a matrix, the matrix/EACA is coated with a protein layer that facilitates EACA's adherence to the matrix. Advantageously, this protein iε thrombin, although other proteinaceous or gelatin compound which facilitates εuch adherence could be utilized, aε well. In a more advantageous embodiment, the matrix is coated with a protein layer prior to application of EACA. In a further advantageous embodiment, the matrix is treated before and after addition of EACA with a protein, preferably which is in solution with an ion additive, εuch aε calcium (i.e., calcium chloride εolution). For example, embodiments such as GT(Ca++)E or CT(Ca++)E, are made by applying to a wound contacting surface of a matrix of gelatin foam or collagen, a firεt solution of thrombin disεolved in calcium chloride, the thrombin preεent at an amount, for example, between 1- 1000 IU/cm 2 , advantageouεly 1-100 IU/cm 2 , and more advantageouεly 1-4 IU/cm 2 , or 1.25 IU/cm 2 . The thrombin is dissolved in 20-60 mM calcium chloride, preferably about 40mM, such that an amount between 25-150 micrograms/cm 2 , preferably 50-100 microgramε/cm 2 , iε depoεited onto that surface. The next step comprises applying to the thrombin-coated matrix surface, 10-100 mg/cm 2 of epsilon aminocaproic acid, preferably 60-70 mg/cm 2 , and preferably in a powder form; then, applying a second solution of thrombin in calcium chloride, which, for example contains the amounts of thrombin and calcium as described in the first solution; and then drying the thrombin, calcium chloride and epsilon aminocaproic acid on the patch. The amount of thrombin applied in the firεt and second solutions can vary, or, a single thrombin solution sealing step can be applied after addition of EACA. Preferably, the total amount of thrombin applied to the wound-contacting surface of the patch by the two stepε iε 2-10 IU/cm 2 . The drying εtep iε accomplished by lyophilization, preferably. Other drying procedures appropriate for a material containing an active protein ingredient can also be employed, so long aε the drying treatment does not denature the proteins or render them inactive when exposed to animal blood. Alternatively, the patch is conventionally dried, by maintaining it at room temperature for a period of 1-3 hours, followed by refrigeration overnight. In other embodiments, for example, GT(Ca++)EF or CT(Ca++)EF, an amount of fibrinogen effective for accelerating hemostaεis in the local environment of the patch. Preferably, an amount of fibrinogen between 2-10 mg/cm 2 is added aε an agent to a matrix which additionally containε EACA, thrombin, and calcium chloride. In yet another embodiment, an agent added to a matrix, in addition to EACA, thrombin, calcium chloride, and optionally, fibrinogen, includeε an amount of protamine sulfate effective to neutralize heparin in the local environment of the patch. Protamine sulfate is added in an amount between 1-15 mg/cm 2 of said matrix, preferably in an amount between 2-5 mg/cm 2 of a wound contacting surface of the matrix. Likewise, RGD or RGDS peptide can be dissolved in double distilled water and sprayed onto a wound- contacting surface of the patch. A patch advantageously contains an amount of RGD effective to enhance clot formation. RGD or RGDS iε applied to a patch advantageously in an amount between 110-130 mg/cm 2 . Thus, a standard size patch would contain about 1-10 mg/patch or about 5-7 mg/patch of RGD or RGDS. It should be noted that, like EACA, the hemostatic agents or additives described in the foregoing paragraphs can be applied to a matrix as a layer, for example, by spraying them onto the wound-contacting surface of the matrix in dried forms. Alternatively, a matrix can be dipped or coated with a solution containing the hemostatic agent/additive. It is desirable that the matrix and agents commingle, particularly when the patch iε exposed to a body fluid such as blood, which permits the dried agents to solubilize and mix. Thuε, a patch can be provided wherein the hemoεtatic agent or mixture of hemostatic agents are absorbed into the pores or interstices of the matrix, or, the agents can be layered on a surface of the matrix and upon solubilizing the agents by addition of body fluid, the desired commingling is achieved. The matrix can be coated with appropriate hemostatic agents described in the above embodiments on one or all surfaces. In a preferred embodiment, the hemostatic agents and additives are coated on only one surface (the wound-contacting surface) . Such an arrangement avoids inducing hemostaεis between the wound and a non-wounded tissue in the vicinity of the patch. In an embodiment intended for packing a void in body tissue, for example, the patch is coated with hemostatic agent(s) /additive(ε) on all surfaces. A kit according to the invention comprises any of the above described hemostatic patch embodiments (which vary in ways including hemoεtatic agent(ε) and additive(ε) utilized, εhape or size) according to the invention and a package, wherein the patch iε contained within a εealed εterile package which facilitates removal of the patch without contamination. The kit can contain multiple patches, preferably wherein each patch is contained within a separate sealed sterile package. A kit designed for field/military use can, in addition to a hemoεtatic patch, further include diεpoεable pre-sterilized surgical instrumentε, εuch as a scalpel, clamp, tourniquet, elastic or inelastic bandage, or the like. Another type of kit comprises a patch containing agents added to the matrix including thrombin, EACA calcium chloride, and protamine sulfate. Such a kit can be prescribed, for example, to patients requiring anticoagulant therapy, to avert the risk of seriouε bleeding which can occur from a minor injury. The availability of such a patch can reduce postoperative hospitalization for patients on dicumarol who underwent surgery. The present invention is further described with reference to the following, illuεtrative exampleε. Unless defined otherwise, all technical and εcientific terms used herein have the same meaning aε commonly understood by one of ordinary skill in the art of the invention. Although any methods and materials similar or equivalent to those described herein can be used in the practice of the invention, the preferred methods and materials have been described. Unless mentioned otherwise, the techniqueε employed or contemplated herein are εtandard methodologieε well known to one of ordinary skill in the art. The materials, methods and examples are illustrative only and not limiting. EXAMPLE 1: THE EFFECTS OF EACA ON THROMBIN ACTIVATION A two-part experiment was designed to teεt whether thrombin activation in the preεence of EACA (A) iε accelerated and (B) is pH dependent. A. Effect of Time Incubated at 37° C. The first part of this study examined activation of thrombin and its degradation in H 2 0 after incubation at 37°C. The assay used was a colorimetric cleaving of a tripeptide, TFA-phe-pro-arg-AFC, where the AFC is the colorometric tag. Seventeen mg of this substrate was dissolved in 200μl DMSO. Thrombin was made up as 10 unitε/ml. The "TEST" εolution contained lOOμl substrate and 200μl of the thrombin solution; a blank contained the same amount of substrate and 200μl of H 2 0. Figure 1 labeled as "ACTIVATION OF THROMBIN SOLUTION AT 37°C" shows the results of that experiment. The optical density in all of these experiments iε an indication of the color and therefore the amount of cleavage of the enzyme that has taken place. The slope of the black-box line indicates that thrombin activation of thrombin dissolved in H 2 0 takes place slowly over a 172 minute time period. The blank, containing substrate and H 2 0, shows no change in optical density, indicating that no activation, or cleaving of the peptide has occurred. B. Thrombin Activation by EACA: A pH Effect In thiε experiment, the hypotheεiε that the activation of thrombin by EACA waε due to EACA's effect of increasing pH was tested. All solutions were prepared at the same concentration as indicated in part A above, except EACA which was made up at a concentration of 50mg/ml. The following samples were prepared: 1. 50μl Thrombin + 925μl H 2 0 + 25μl subεtrate 2. 50μl Thrombin + 925μl Tris buffer @ pH 7.02 + 25μl substrate 3. 50μl Thrombin + 925μl Tris buffer @ pH 7.62 + 25μl subεtrate 4. 50μl Thrombin + 925μl Triε buffer @ pH 7.80 + 25μl substrate 5. 50μl Thrombin + 925μl Tris buffer @ pH 8.01 + 25μl substrate 6. 50μl Thrombin + 425μl EACA sol. + 500μl H 2 0 + 25μl substrate 7. 50μl Thrombin + 425μl EACA sol. + 500μl Triε buffer @ pH 7.02 + 25μl εubstrate 8. 50μl Thrombin + 425μl EACA sol. + 500μl Tris buffer @ pH 7.62 + 25μl subεtrate 9. 50μl Thrombin + 425μl EACA sol. + 500μl Tris buffer @ Ph 7.80 + 25μl substrate 10. 50μl Thrombin + 425μl EACA sol. + 500μl Tris buffer @ Ph 8.01 + 25μl substrate Each tube was placed in a 37°C. waterbath and removed periodically to be read each 5' for a total of 60'. Results are summarized in Figure 2. In the legend, samples 1-10 listed vertically in the legend correspond to samples 1-10 immediately above, while "T" representε thrombin. The results indicate clearly that the action of EACA is a pH effect and that Tris buffer-adjusted solutions had a similar effect aε the pH waε increaεed. In all caεeε, the plateau may not be accurate εince the εaturation of the inεtrument occurs near to the maximum optical density recorded. At 37°C, the reεultε indicated clearly that the action of EACA iε a pH effect. Calcium ion appears to enhance this pH-mediated activation. EXAMPLE 2: EACA EXERTS AN ANTIBACTERIAL EFFECT EACA was shown to inhibit both Staph . aureus and E . coli in a doεe-dependent manner by the following method. Culture plateε and EACA diεcs were prepared as follows: Whatman filter paper discs of 5.4 cm in diameter and 22.9cm 2 in area were placed in beakers of almost the same diameter. EACA (229mg) waε disεolved in 250μl of double diεtilled H 2 0 and used to make the final concentrations. All concentrations of EACA were applied in 250μl of H 2 0. Concentrations of 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100 mg/cm 2 were prepared. After application of EACA solutionε, the discs were allowed to dry and frozen to ensure stability. Discs for application to agar plates were made with a paper punch, at a size of about 6.35mm. Agar plates were poured in two increments. A first increment of 1.5% Brain-Heart Infusion agar waε prepared and autoclaved. After cooling to approximately 55°C, 12 mlε were added to each 100mm x 15mm petri diεh. Plateε were allowed to cool to room temperature, wrapped in parafilm and refrigerated. Brain- Heart Infusion broth was prepared and autoclaved. When the temperature was cooled to room temperature, a 1ml aliquot of Staph . aureus or E . coli was added and the broth incubated overnight at 37°C. The following day, a second increment of 1.2% Brain- Heart Infusion agar was prepared and when cooled to 48°C after autoclaving, 2ml of each culture was added to separate flaskε of agar and 1ml of these mixtures waε added to each culture plate. This top layer was allowed to harden at room temperature. Two setε of five diεcs containing EACA at varying concentrations were added to each plate, in addition to a control disc containing zero mg/cm 2 EACA. The complete results are listed in Table 2. Figure 3A and Figure 3B each show inhibition by EACA of Staph . aureus growth graphically, for each set of various concentrations of EACA, while Figures 4A and Figure 4B each show inhibition by EACA of E . coli growth for each set of varying concentrations of EACA. Reεults of observation and meaεurement of the zone inhibition reveal that in almost all instances, there is an incremental change in this zone of inhibition related to the concentration of EACA. The exceptions are that the 60mg/cm 2 did not follow the trend, but was equal to or decreased in relation to the 40mg/cm 2 . The 90 and lOOmg/cm 2 zoneε were not always increases. The consiεtency of theεe variations appear to be related to the disc preparation rather than a biological variation. TABLE 2. Results: Inhibition of Staph . aureus (Plates 1-6) and E . coli (Plates 7-12) Growth by EACA Date Plate Organism Cone, of DIAMETER % > % OF Number EACA OF CONTROL MAXIMUM in INHIBITION mg/cm2 10/22/93 1 E. coli control 6.35 0.00 77.00 10 6.95 9.40 84.20 30 7.65 20.50 92.70 50 7.75 22.00 93.90 70 8.25 29.90 100.00 90 7.50 18.10 90.90 10/22/93 2 E. coli control 6.35 0.00 70.20 10 6.70 5.50 74.00 30 8.55 34.60 94.50 50 8.60 35.40 95.00 70 9.05 42.50 100.00 90 8.35 31.50 92.30 10/22/93 3 E. coli control 6.35 0.00 70.60 10 6.70 5.50 74.40 30 7.05 11.00 78.30 50 7.10 11.80 78.80 70 9.00 41.70 100.00 90 8.25 29.90 91.70 10/22/93 4 E. coli control 6.35 0.00 77.40 Date Plate Organism Cone, of DIAMETER % > % OF Number EACA OF CONTROL MAXIMUM in INHIBITION mg/cm2 20 7.05 11.00 86.00 40 7.70 21.30 93.90 60 7.70 21.30 93.90 80 8.20 29.10 100.00 100 7.75 22.00 94.50 /22/93 5 E. coli control 6.35 0.00 78.40 20 7.70 21.30 95.10 40 7.75 22.00 95.70 60 7.45 17.30 92.00 80 8.10 27.60 100.00 100 8.10 27.60 100.00 /22/93 6 E. coli control 6.35 0.00 76.50 20 7.60 19.70 91.60 40 8.10 27.60 97.60 60 7.90 24.40 95.20 80 8.25 29.90 99.40 100 8.30 30.70 100.00 /22/93 7 S. aureus control 6.60 0.00 77.20 10 7.55 14.40 88.30 30 8.20 24.20 95.90 50 7.65 15.90 89.50 70 8.55 29.50 100.00 90 7.90 19.70 92.40 Date Plate Organism Cone, of DIAMETER % > % OF Number EACA OF CONTROL MAXIMUM in INHIBITION mg/cm2 /22/93 8 S. aureus control 6.55 0.00 80.90 10 7.10 8.40 87.70 30 7.00 6.90 86.40 50 7.15 9.20 88.30 70 7.75 18.30 95.70 90 8.10 23.70 100.00 /22/93 9 S. aureus control 6.55 0.00 81.90 10 7.00 6.90 87.50 30 7.20 9.90 90.00 50 7.45 13.70 93.10 70 7.85 19.80 98.10 90 8.00 22.10 100.00 /22/93 10 S. aureus control 6.60 0.00 79.00 20 8.05 21.90 96.40 40 8.30 25.80 99.40 60 8.10 22.70 97.00 80 7.55 14.40 90.40 100 8.35 26.50 100.00 /22/93 11 S. aureus control 6.50 0.00 83.90 20 7.20 10.80 92.90 40 7.70 18.50 99.40 60 7.30 12.30 94.20 80 7.40 13.80 95.50 100 7.75 19.20 100.00 0/22/93 12 S. aureus control 6.50 0.00 79.30 20 6.75 3.80 82.30 40 8.05 23.80 98.20 60 8.00 23.10 97.60 80 8.20 26.20 100.00 100 7.20 10.80 87.80 EXAMPLE 3: A COMPARISON BETWEEN G(Ca++)TE AND TACHOCOMB® A. Experimental Conditions 1. Patch Preparation An absorbable gelatin sponge, namely a gelatin foam matrix (GelFoam®, UpJohn Co.) was obtained. Physician ' s Desk Reference 2451, 47th Edition Dowd (ed.), Medical Economics Data (1993) . Thereafter, 1.25 IU/cm 2 bovine thrombin was applied to a surface of the gelatin foam. Next, either 10mg/cm 2 or lOOmg/cm 2 of EACA waε applied to that same surface, followed by an application of another 1.25 IU/cm 2 application of bovine thrombin. The patcheε were allowed to dry and left in a refrigerator overnight. A "blank" gelatin foam patch, which waε not treated with thrombin or EACA was also tested in the kidney. TachoComb® patches were obtained and applied according to the manufacturer's inεtructions. That is, prior to preparation, the TachoComb® patches were dipped in sterile saline and applied to bleeding organs with light pressure for five minutes. 2. Organ Preparation A lobe of pig liver waε εurgically iεolated and three lesions approximately 1x1.5 cm in size were created. Blood flowed freely from each of the lesions. Each of the patches discussed in part A. (above) , were applied and kept under pressure by a saline soaked sponge for five minutes and the pressure was released. Patches were evaluated by their ability to control hemorrhage in termε of (a) leakage, (b) ability to withεtand increaεed vascular pressure, (c) the resiεtance offered when attempting to peel the patch from the lesion, and (d) events of clot formation in the lesion. For the liver, pressure tests were performed by raising the arterial presεure by injecting 0.2 ml 1\1000 epinephrine. For renal studies, both poles (ends) of the kidney were surgically removed to a depth of approximately 0.5 cm, while the renal artery was clamped. The clamp was removed after the test patches were placed and presεure applied with a εaline εoaked εurgical εponge for five minuteε. B . Summary of Results In liver, when the preεεure was removed and after five minutes, both patches according to the invention showed good control of hemorrhage, with only a little bleeding from the edge in the lOOmg patch and no bleeding from the lOmg patch. After 9-13 minutes, the TachoComb® patch waε the only patch leaking or bleeding from the edge. Reεultε are εhown in Figureε 6 and 7, which show a Hemarrest™ patch containing either 100 mg/cm 2 (left side) or 10 mg/cm 2 (right side) epsilon aminocaproic acid, respectively. In Figure 6, blood iε preεent along the lower edge of the TachoComb® patch and between that patch and the 100 mg/cm 2 patch. Thiε bleeding originated from the TachoComb® patch. A small amount of blood is present on the surface of the lOOmg patch, while none is present on the lOmg patch. Figure 7 showε the patches removed from the same liver shown in Figure 6. Free blood is present coming from the lOOmg and TachoComb® lesionε. A greater flow is observed coming from the TachoComb® patch. Much of the clot from the TachoComb® site stays with the patch when it is peeled back. A piece of the gelatin foam patch iε incorporated into the lOmg εite. When epinephrine waε injected, the TachoComb® patch still dripped blood from the edges after 18 minuteε. The peel teεt after 20 minutes showed the TachoComb® patch with minimal adheεion, the clot stuck to the patch, and the wound continued to bleed. In the lesion with the lOOmg patch, blood also flowed, but not aε much as the TachoComb® patch. The lOmg patch had the least bleeding of any of the patches and had both good incorporation of the patch into the lesion and good clot formation, with some minimal adhesion to the periphery. In the kidney, there was not much difference between the TachoComb® and the lOOmg patch lesions. There was no bleeding before or after epinephrine injections. When the patches were peeled at 20 minutes, the TachoComb® patch had very good adhesive qualities, good clot formation, but some free blood. The lOOmg patch did not have as good adhesiveneεs, but had a well-formed clot and no hemorrhage. When a blank gelatin patch and lOmg patches according to the invention were compared, the lOmg patch definitely was better. Five minutes after the pressure release, there was free blood under the lOOmg patch while there waε some bleeding around the edge of the lOmg patch. This waε unchanged after epinephrine, but when an experimental peel test was done by removing the patch and observing clot formation, the clot was not aε well-formed under the blank patch. Further, free blood waε preεent, and there waε a blot stain dark with blood on the dry surgical sponge held against the patch to detect blood or serum penetrating the patch. There was good adhesion of the lOOmg patch to the surface even when the patch is removed. The lOmg patch had fair adhesion around the edges and some free blood. When the patch was lifted there waε evidence of good clot formation and no bleeding, thereby providing a light pink blot test measured by the dry εurgical sponge held against the patch. Figure 8 illustrates a patch applied to a kidney, the patch containing 2.5 IU/cm 2 thrombin and lOOmg/cm 2 epsilon aminocaproic acid. The light pink color indicateε that virtually no free blood penetrateε through the patch. No blood iε present on the sponge that held the hemostatic patch against the organ. Figure 9 shows the opposite pole of the same kidney of Figure 8, covered with a TachoComb® patch. The latter patch is darker, which indicates that more blood is comes through the patch matrix. The lower edges of that patch are looser as compared to the patch of Figure 7. Fresh blood could be seen on a dry sponge held against the organ for the purpose of aiding in detection of fresh blood. EXAMPLE 4: HEMOSTATIC EFFICACY ACHIEVED BY THE GE(Ca++) PATCH 1) Pig splenic lesionε were created as discuεsed in Example 3. As εeen in Figure 10, no leakage waε obεerved from the GE(Ca++) patch, while εome waε observed from the GT(Ca++)E patch. In 10 minutes, there was slight leakage from the centers of both, which waε εtopped by 15 minuteε. When the patcheε were removed, there was no difference in a test blotting performed on the εurface of the patch, as both test blots were light pink. Very good adhesion was observed for both patches, as well as large, well-formed clots. In the GE(Ca++) patch, the clot adhered to the patch but not the lesion. 2) In the liver, neither showed bleeding at any observation. When peeled, the patcheε both had good adhesion, but the GE(Ca++) patch bled freely after the patch was removed. In contrast, the GT(Ca++)E patch had some incorporation and a good clot. The GE(Ca++) did not seem to have a good clot. 3) The kidneys had unexpected findings. The GE(Ca++) patch had no evident leakage while the GT(Ca++)E leaked steadily. At 10 minuteε, the leakage had leεεened in the GT(Ca++)E patch, and at 15 minuteε, there waε no further leakage in either. When the patches were removed, both had good adhesion, εome incorporation of the GT(Ca++)E patch, but both bled in the abεence of the patch. The concluεion from thiε one experiment εuggests that there iε little difference between the treatmentε although clot formation appears to be better with the addition of thrombin. This means that a first-aid bandage that is εtable under more εevere expoεure to heat may be effective without the presence of thrombin. EXAMPLE 5: A COMPARISON OF CVA(Ca++)TE ("CALCIUM/SODIUM ALGINATE") AND OTHER PATCHES 1) Patches CVAT(Ca++)EF, CVCT(Ca++)EF and GT(CA++)EF, were applied in the usual manner to leεionε on the εpleen and each contained 2.5mg/cm 2 of fibrinogen. Aε shown in Figure 11, the resultε demonstrate that GT(Ca++)EF, CVAT(Ca++)EF, and CVC(Ca++)EF bled from the edge at release of presεure at 5 minuteε. At 10 minuteε, the bleeding stopped except for the uncovered part of the lesion. At 15 minutes, all hemorrhage has stopped. When the patches were removed: GT(Ca++)EF began to bleed when the lesion was stretched but had a well-formed clot and good adhesion within the wound; CVAT(Ca++)EF had good adheεion within the wound, εome clot formation and free blood under the patch; and CVC(Ca++)EF had better adheεion, and a well formed clot in the leεion, thiε was the most effective of the three patcheε. An evaluation approximately 50 minuteε later showed that (1) CVAT(Ca++)EF had the best adhesion, (2) GT(Ca++)EF adhered to the lesion, and (3) CVCT(Ca++)EF was loose but not bleeding either. Similar studies were performed in liver and kidney, with the conclusion being drawn that there was not much difference between the patcheε teεted, although CVAT(Ca++)EF slightly outperformed the other two patches. EXAMPLE 6: THE GT(Ca++)EFP ("PROTAMINE") PATCH This study was performed to test protamine, a heparin antagonist, in the combination of compounds applied to the patches. Protamine iε advantageouε to facilitate patientε who are anticoagulated for myocardial, pulmonary, vaεcular disease or other disease in which prevention or prolongation of coagulation iε of advantage. To test the protamine patch, a pig was heparinized at the dose of 2000 units before the study began and another 1000 units one hour later and 45 minutes before termination of the study. The patches GT(Ca++)EF "Gelfoam", CT(Ca++)EF "Collagen", and GT(Ca++)EFP ("Protamine") uεed in thiε εtudy are εhown in Figure 12A. 2 Protamine waε applied to the Protamine Patch in an amount of 5 mg/cm 2 . Fibrinogen and EACA were ground to a fine powder before application to the patches in amounts, as described previously. 1) In the spleen, all lesionε continued bleeding after the preεεure waε releaεed, but the leεion with a protamine patch bled the leaεt. The photo shown in Figure 12B waε taken 35 εecond after releaεe of preεεure. Preεsure waε reapplied from 7.5 to 10 minuteε. At 10 minuteε the pressure was again removed and bleeding aεεeεεed. All leεions continued to bleed. Pressure was applied once again for about 4 more minutes. Now, GT(Ca++)EF had controlled the hemorrhage; CT(Ca++)EF was bleeding only from an exposed part of the lesion where the patch had slipped; and, GT(Ca++)EFP waε similar to CT(Ca++)EF. Pressure waε reapplied for 45 seconds. Patches were removed from the lesionε 3 minuteε later. GT(Ca++)EF had no bleeding, good incorporation and adheεion, but little clot; CT(Ca++)EF waε εtill bleeding from the exposed part of the lesion, had good adhesion and a good clot, but waε still bleeding under the patch; and, GT(Ca++)EFP continued to bleed from the exposed portion of the lesion, had good adhesion, incorporation, and clot formation. The spleen was ligated. The abdominal cavity had a large clot from hemorrhaged blood. 2) The protocol was similar for the liver. The GT(Ca++)EF patch waε bleeding heavily from the edgeε; the CT(Ca++)EF waε bleeding from the edges; and, the GT(Ca++)EFP had only slight bleeding from the edges. Preεsure was reapplied for 15 seconds and then none of the patches showed hemorrhage at the 10 minute mark. This waε true at 15 minutes, as well. At 20 minutes, the patcheε were peeled off. GT(Ca++)EF had good adheεion but free blood under the patch; CT(Ca++)EF had less adhesion but good clot formation; and GT(Ca++)EFP had both good adhesion and clot formation under the patch. 3) In the kidney, the results were similar. After presεure release, the GT(Ca++)EF patch was leaking in the center; CT(Ca++)EF was oozing from the bottom of the patch; and, GT(Ca++)EFP was oozing from the middle and soaked in appearance. At 10 minutes, GT(Ca++)EF was leaking slightly if any; and, CT(Ca++)EF and GT(Ca++)EFP had no bleeding. Epinephrine was injected. GT(Ca++)EF began to leak; CT(Ca++)EF began to leak from the edge; and, GT(Ca++)EFP did not leak. When the patches were removed, GT(Ca++)EF had fair adhesion, some free blood and oozing from the surface; CT(Ca++)EF had very little adhesion, more free blood, but good clot formation; and, GT(Ca++)EFP had fair adhesion, a good clot and no hemorrhage. In summary, a comparison was made between gelatin foam and collagen patches prepared with thrombin, EACA, and fibrinogen, with a gelatin foam patch containing the same compounds plus protamine, a heparin antagonist. While it took longer to effect hemostasiε, pressure had to be applied several times between observations and bleeding control took longer, the resultε of all experimentε indicated that addition of protamine εulfate resulted in earlier stoppage of blood flow, better clot formation and usually better adhesion than those patches without protamine sulfate. The results predict that when a patient is heparinized or receiving dicumarol, it is advisable to apply pressure to a protamine patch for aε long aε 20 minutes, and preferably at least 10 minutes. EXAMPLE 7: THE ("RGD") PATCH The study has both partε I and II. Part 1: Patches CTR, CTE(f) , GT(f)E(f) and a plain gelfoam (G) patch were applied to leεionε made on the spleen of an anesthetized pig. The symbol "(f)" denotes the compound immediately preceding it aε a £reshly- applied compound. That iε, E(f) denoteε EACA that is freεhly applied to a patch very soon (lesε than about three hourε) after it is made. 1) Leakage : When the sponge preεεure waε removed from the patcheε, the G patch had virtually no leakage. Thiε waε true of the CTE(f)R patch aε well, but the CTR patch εhowed much bleeding. Shortly thereafter, the reεults were recorded as εimilar. 2) Peel /Adhesion : All three patches stuck to the saline-soaked sponges and removal of sponge pressure was done carefully to prevent their removal from the lesion; thus adhesion in all patches was minimal at that time. Patch G did show some adhesion, but CTR and CTE(f)R εhowed good adheεion even though they each had εome clot formation, beεt in CTE(f)R. Approximately 6 minutes after removing the pressure, the gelatin foam showed very good adhesiveness and poor clot formation. Neither of the other patches showed good adhesion qualities, while the CTR some clot and CTE(f)R had a large, excellent clot. Part II: More lesions were created on the spleen and all reεults were compared. Patcheε applied were CTE(f) and GT(f)E(f) . 1) Leakage : Neither the CTE nor the GT(f)E(f) patch showed leakage at removal of sponge presεure. Five minuteε later, the vein waε occluded and intravaεcular preεsure increased and lesions made in both parts were evaluated. The time for the increaεed preεεure teεt performed after the εponge waε released in Part I is 27 minutes and in Part II, only 5 minutes. The gelatin foam only patch (G) was not leaking at all; neither were CTR, ** CTE(f)R. In comparing the two patches with T and EACA using either the gelatin foam or collagen matrix, the gelatin foam patch, GT(f)E(f), showed lesε leakage than the collagen based patch, CTE. In fact, the CTE patch leaked more as the venous pressure was raised. 2) Peel /Adhesion : Two minutes after removing the sponge presεure and when comparing CTE(f) , collagen + T + EACA and GT(f)E(f) , the gelatin foam baεed patch similarly treated showed very good adhesion to both the lesion and the surrounding tissue. The CTE patch with the collagen base had little or no adhesion. 10 minutes after removing sponge pressure (part II patches GT(f)E(f) and CTE(f)) and with the intravascular pressure still elevated, all patches from Part I and Part II were evaluated together. The time interval was about 32 minutes after initial sponge pressure removal for those patches from Part I (CTR, G, and CTE(f)R) . The results were as follows: Patch CTR) No adhesion, good clot formation and little leakage. Patch G) Strong adhesion to surrounding tissue, no adhesion to lesion, much leaking. Patch CTE(f)R) No adhesion, good clot formation, little leakage. Patch GT(f)E(f)) Good adhesion to lesion, good clot formation, little leakage. Patch CTE(f) No adhesion, good clot formation, little leakage. 3) A further asεessment of εplenic leεionε waε made (Part I) 58 minuteε after initial sponge pressure release, Part II, -36 minutes after initial pressure release. The patcheε were removed at this time. The results of this assessment are: Patch TR) Moderate clot formation, little, if any, leakage. Patch G) Leakage, but remainder of gelatin foam stuck to lesion. Patch CTE(f)R) Excellent clot, no leakage. Patch GT(f)E(f)) Good clot formation, little, if any, leakage. Patch CTE(f) good clot formation, virtually dry. 4) A final assessment of these splenic lesions was made 37 minutes later. Patch CTR) Dry to the blot teεt (placing a dry surgical sponge on the lesion) , clot is developed, no collagen incorporated into lesion. Patch G) Dry to the blot test, gelatin foam incorporated into the lesion. Patch CTE(f)R) Absolutely no blood elements on sponge after blotting; clot is excellent, filling lesion and extending onto the surrounding normal area. Patch GT(f)E(f)) Serum staining on sponge, but good clot and gelatin sponge incorporated into the lesion. Patch CTE(f) Dry to blot test; similar to CTE(f)R.

Claims: We Claim: 1. A hemostatic patch comprising a structural element that iε a biodegradable matrix having a wound- contacting εurface, wherein εaid matrix iε εelected from the group consisting of absorbable gelatin sponge, calcium alginate, calcium/sodium alginate, collagen, and oxidized regenerated celluloεe, to which matrix iε applied a hemostatic agent comprising an amount of epsilon aminocaproic acid effective for inhibiting fibrinolysis. 2. A hemostatic patch according to claim 1, wherein the matrix is absorbable gelatin sponge. 3. A hemostatic patch according to claim 2, wherein said absorbable gelatin εponge is gelatin foam. 4. A hemostatic patch according to claim 3, wherein said gelatin foam is compressed to at least about one- half its original thickness. 5. A hemostatic patch according to claim 1, wherein the matrix is collagen. 6. A hemostatic patch according to claim 1, wherein the matrix is calcium alginate. 7. A hemostatic patch according to claim 1, wherein the epsilon aminocaproic acid is present in an amount between 10-100 mg/cm 2 of the wound-contacting surface of the matrix. 8. A hemostatic patch according to claim 4, wherein the epsilon aminocaproic acid is preεent in an amount between 10-100 mg/cm 2 of the wound-contacting surface of the matrix. 9. A hemostatic patch according to claim 1, wherein said matrix further comprises an amount of thrombin effective for promoting accelerated hemostaεiε. 10. A hemoεtatic patch according to claim 9, wherein the thrombin iε bovine thrombin. 11. A hemostatic patch according to claim 9, wherein the thrombin is present in an amount between 1-1000 IU/cm 2 of the wound-contacting surface. 12. A hemostatic patch according to claim 11, wherein the thrombin is present in an amount between 2-10 IU/cm 2 of the wound-contacting surface. 13. A hemostatic patch according to claim 9, wherein the matrix further comprises calcium ions in an amount effective for stimulating hemostasis. 14. A hemostatic patch according to claim 9, wherein calcium chloride is present in an amount between 25-150 micrograms/cm 2 of the wound-contacting surface. 15. A hemostatic patch according to claim 14, wherein the amount of calcium chloride is between 50-100 micrograms/cm 2 . 16. A hemostatic patch according to claim 13, the patch further comprising an amount of fibrinogen effective for stimulating hemostasis. 17. A hemostatic patch according to claim 16, wherein the fibrinogen is bovine fibrinogen. 18. A hemostatic patch according to claim 17, wherein the fibrinogen is present in an amount between 2- 10 mg/cm 2 of the wound-contacting surface. 19. A hemostatic patch according to claim 13, which further comprises an amount of RGD peptide effective for accelerating hemostasiε. 20. A hemostatic patch according to claim 19, wherein RGD is present in an amount between 10-1000 mg/cm 2 of the wound-contacting surface. 21. A hemostatic patch according to claim 20, wherein said amount of RGD is 50-150 mg/cm 2 . 22. A hemostatic patch according to claim 19, wherein said RGD peptide further comprises serine. 23. A hemostatic patch according to claim 13, which further comprises an amount of protamine sulfate effective for neutralizing heparin present in the local environment of the patch. 24. A hemostatic patch according to claim 19, wherein protamine sulfate is present in an amount between 1-15 mg/cm 2 of the wound-contacting surface. 25. A hemostatic patch according to claim 24, wherein protamine sulfate is present in an amount between 2-5 mg/cm 2 . 26. A hemostatic patch according to claim 1, the patch having a thickness of 2-10 millimeters. 27. A method for effecting hemostasiε in a bleeding wound, compriεing applying a wound-contacting surface of a hemostatic patch according to claim 1 to a bleeding wound, and maintaining the patch in contact with the wound for a period of time εufficient to permit clotting to occur at the interface between the hemostatic patch and the wound, and for bleeding to be substantially arrested. 28. A method according to claim 27, wherein said period of time is between 3 to 20 minutes. 29. A method according to claim 28, wherein said period of time iε between 3 to 5 minuteε. 30. A method according to claim 27, further compriεing the εtep of moiεtening the patch with a εterile εaline εolution prior to applying the patch to a wound. 31. A kit compriεing a sterile package containing a patch according to claim 13. 32. A kit according to claim 31, further compriεing εterilized surgical instruments. 33. A kit according to claim 32, further comprising at least one bandage. 34. In a method for effecting hemostasis by applying a hemostatic patch containing one or more hemostatic agents to a wound, the improvement wherein said patch comprises a composition capable of raising the pH at the site of application of the patch to a pH in the range of 7-9, whereby the activation of thrombin at said site iε accelerated. 35. A method according to claim 34, wherein εaid compoεition iε epεilon aminocaproic acid. 36. A method according to claim 34, wherein εaid compoεition comprises a buffer for maintaining the pH between 7-9. 37. A hemostatic patch which is made by a process comprising the steps of: (a) applying a solution of bovine thrombin, dissolved in 30-50 mM calcium chloride, to a wound- contacting surface of a structural element that is a biodegradable matrix of compressed gelatin foam; (b) applying epsilon aminocaproic acid to said wound-contacting surface in an amount between 1-100 IU/cm 2 of said surface; (c) applying a solution of bovine thrombin, dissolved in 30-50 mM calcium chloride, to seal the surface, such that the total amount of bovine thrombin present is between 1-1000 IU/cm2 of said surface; and (d) drying the resultant patch. 38. A hemostatic patch comprising a flat, flexible matrix that is biodegradable and absorbable, having an uncompressed thickness of about 4-10 mm and a length, and applied to a single wound-contacting surface along the length of said matrix, EACA in amounts 10-100 mg/cm 2 and optionally applied to said surface, one or more additives, including a calcium ion source, RGD peptide, RGDS peptide, protamine sulfate and buffer. 39. A patch according to claim 38, further comprising about 1-1000 IU/cm 2 of thrombin applied to said surface. 40. A hemostatic bandage for applying to a wound on a patient, said bandage comprising a backing member fixedly secured to a patch according to claim 38; and at least one flap extending from said backing member beyond said patch, said flap having applied thereto a medically acceptable adhesive suitable for adhering the bandage to the patient.

Inventors: PRUSS THADDEUS P, WILL JAMES A, PRUSS, THADDEUS, P., WILL, JAMES, A.
Applicants: CLARION PHARMACEUTICALS INC, PRUSS THADDEUS P, WILL JAMES A, CLARION PHARMACEUTICALS, INC., PRUSS, THADDEUS, P., WILL, JAMES, A.
Application year: 1994
Main classification: A61F13
IPC codes: A61F13, A61L15, A61L24, A61L26